| Object:To investigate the changes of expression of Neurotrophin-3(NT-3) and the synaptic density in the dentate gyrus of hippocampus temporal lobe Epilepsy rats,and to determine the potential significance of the changes for the development of temporal lobe Epilepsy.Method:Control(16rats)and experimental(24 rats)groups were set up. Temporal Lobe Epilepsy molds were established in adult rats through lateral cerebral ventricle injection of kainic acid for experimental group.6h,24h,48h,7d after injection,cut the head and took the brain tissue.The normal sodium took the place of KA in control group.Investigated the The changes of NT-3 expression and synaptic density after epilepsy in dentate gyrus of the rat's hippocampus by immunohistochemistry methods and immune electron microscope.The t test of two independent samples was used in the comparison between experimental and control groups in the same time point.The analysis of variance(ANOVA)for completely randomized design was used to compare the variance in different time points of the same groups.Significance level a=0.05Result(1)the change of The expression of NT-3 by immunohistochemistry method:compared with the control group,the number of NT-3-positive cells in dentate gyrus in all the experiment groups(6h,24h,48h and 7d)after epilepsy decreased(p＜0.01),(the data of experimental groups in different time points were 192.00±27.70,89.67±14.21,47.33±8.99,113.17±21.28,data of control groups were 254.60±44.90,238.80±12.03,259.40±10.21,241.8±19.90) (p＜0.01).The comparison among experiment groups was used the analysis of variance for completely randomized design:the number of positive cells of 24h group was less than 6h group(p＜0.01),48h group was less than 24h group (p＜0.01),7d group were more than 48h group(p＜0.01).The number of NT-3 positive cells was in the tendency of decrease,it was in the lowest point in 48h interval,increased in 7d interval.There was not difference in the comparison of different time point groups of control groups(p＞0.05).(2)The antibody was marked by golden granule in immune electron microscope experiment.The NT-3 can be localized precisely.In the experiment,the NT-3 marked golden granules were observed in kytoplasm,chondriosomes,axons,dendrites,anterior and posterior mambranes of synapse.Compared with the control group,the number of NT-3-marked golden granules in dentate gyrus in all the experiment groups(6h,24h,48h and 7d)after epilepsy decreased(p＜0.01)(the data of experimental groups in different time points were 9.08±1.58,4.89±0.72,2.89±0.40,6.84±0.80,data of control groups were 16.49±1.87,16.28±1.62,16.61±2.11,16.50±2.40).The comparison among experiment groups was used the analysis of variance for completely randomized design:the number of golden granules of 24h group was less than 6h group(p＜0.01),48h group was less than 24h group(p＜0.01),Group 7d were more than Group 48h(p＜0.01).The number of NT-3 marked golden granules were in the tendency of decrease,it was in the lowest point in 48h interval,increased in 7d interval.There was not difference in the comparison of different time point groups of control groups(p＞0.05).(3)In the experiment groups,the postsynaptic membrane were thicker,the intervals of synapse were broader.The synaptic density of dentate gyrus:compared with control group,there wasn't obvious difference in the synaptic density of Group 6h and Group 7d in experiment groups(p＞0.05),the synaptic density decreased in Group 24h and Group 48h(p＜0.05)(24h:experimental group 3.75±0.58,control group 5.07±0.52;48h:experimental group 3.04±0.46;control group 5.09±0.87).The comparison among experiment groups was used the analysis of variance for completely randomized design:the density of synapse in 24h was less than 6h(p＜0.05),48h was less than 24h(p＜0.05),7d was more than 48h(p＜0.01).The synaptic density decreased first then increased.There was not difference in the comparison of different time point groups of control groups(p＞0.05).(4)There is positive correlation between NT-3 and synaptic density in rats'dentate gyms after epilepsy(r=0.536).Conclusion 1.The quality of NT-3 decreased firstly then growed up,but still less than normal level after epilepsy,the NT-3 marked golden granules were observed in kytoplasm,chondriosomes,axons,dendrites,anterior and posterior mambranes of synapse.2.the process of the synapse density decrease and returned were the process of synapse rebuild and rehabilitation.3.NT-3 was related to the synaptic density,suggested that it participated in the synapse rehabilitation,.
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